Identifying key molecules that mediate brain reward plasticity remains an important goal of current drug abuse research. We recently identified a key role for the Fragile X Mental Retardation Protein (FMRP), a RNA binding protein involved in regulating protein synthesis in dendrites, as an essential downstream component of MEF2-dependent synapse elimination. Moreover, we observed significant deficits in cocaine-induced behavioral plasticity in FMRP-deficient mice. Taken together with the recently documented role for MEF2 in repeated cocaine-induced structural and functional plasticity, our findings in the Fmr1 KO mice suggest an important role for synapse elimination as a process that promotes long-lasting behavioral plasticity associated with chronic cocaine use. In this proposal, we will test the hypothesis that FMRP mediates an acute process of synapse elimination/remodeling in the NAc that is required for behavioral plasticity associated with repeated cocaine exposure. To this end, we propose the following: Specific Aim 1: Our preliminary findings indicate that Fmr1-/- mice have significantly reduced cocaine sensitization and place preference to cocaine. As these are developmental knockout mice, the precise populations of cells where FMRP exerts its function on cocaine-induced behaviors is not clear. To this end, we will take a two-pronged approach: 1) we will express WT FMRP in the nucleus accumbens (NAc) using viral- mediated gene delivery in the Fmr1 KO mice and test for functional rescue of cocaine behaviors, and 2) we will selectively knockout the Fmr1 gene in the NAc using viral-mediated gene delivery of Cre recombinase in adult conditional Fmr1 knockout mice and test the requirement for FMRP in the adult NAc during cocaine-induced behavioral plasticity. Specific Aim 2: Our recent findings revealed that FMRP is strictly required for MEF2-dependent regulation of synapse number in hippocampal pyramidal neurons, and NAc expression of active MEF2 enhances both sensitized locomotion and place preference to cocaine. Therefore, we will test the hypothesis that FMRP functions in the NAc to mediate MEF2-induced sensitized behavioral responses to cocaine. To this end, we will use established viral-mediated gene delivery to determine whether MEF2-modulated behavioral responses to cocaine require FMRP function in vivo using Fmr1-/- mice and established behavioral assays. Specific Aim 3: We previously demonstrated that inhibition of MEF2 activity is necessary and sufficient to regulate the chronic cocaine-induced increase in NAc MSN spine density. Since we found that FMRP is required for MEF2-regulated excitatory synapse density in hippocampal neurons, we will test the hypothesis that the cocaine-induced increase in NAc spine density requires FMRP in vivo. Using established spine imaging methods, we will assess the basal and chronic cocaine-regulated NAc spine density in Fmr1 KO mice. PUBLIC HEALTH RELEVANCE: Drug addiction is a chronic human disease characterized by uncontrolled drug use despite severe adverse consequences to the addict. The results of the proposed studies would provide valuable new insights into molecular and neural circuit mechanisms that contribute to long-lasting behavioral responses to chronic drug use, and could generate new therapeutic targets for the treatment of drug addiction for which there are currently very limited treatment options.